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This study focused on N-acetyltryptamine, a little-studied melatonin receptor mixed agonist-antagonist and the likely evolutionary precursor of melatonin. The central issue addressed was whether N-acetyltryptamine is physiologically present in the circulation. N-acetyltryptamine was detected by LC-MS\/MS in daytime plasma of 3 different mammals in subnanomolar levels (mean \u00b1 SEM: rat, 0.29 \u00b1 0.05 nM, n = 5; rhesus macaque, 0.54 \u00b1 0.24 nM, n = 4; human, 0.03 \u00b1 0.01 nM, n = 32). Analysis of 24-h blood collections from rhesus macaques revealed a nocturnal increase in plasma N-acetyltryptamine (p &lt; 0.001), which varied from 2- to 15-fold over daytime levels among the 4 animals studied. Related RNA sequencing studies indicated that the transcript encoding the tryptamine acetylating enzyme arylalkylamine N-acetyltransferase (AANAT) is expressed at similar levels in the rhesus pineal gland and retina, thereby indicating that either tissue could contribute to circulating N-acetyltryptamine. The evidence that N-acetyltryptamine is a physiological component of mammalian blood and exhibits a daily rhythm, together with known effects as a melatonin receptor mixed agonist-antagonist, shifts the status of N-acetyltryptamine from pharmacological tool to candidate for a physiological role. This provides a new opportunity to extend our understanding of 24-h biology.<\/jats:p>","DOI":"10.1177\/0748730417700458","type":"journal-article","created":{"date-parts":[[2017,5,3]],"date-time":"2017-05-03T06:44:51Z","timestamp":1493793891000},"page":"195-211","update-policy":"https:\/\/doi.org\/10.1177\/sage-journals-update-policy","source":"Crossref","is-referenced-by-count":23,"title":["Daily Rhythm in Plasma N-acetyltryptamine"],"prefix":"10.1177","volume":"32","author":[{"given":"Peter S.","family":"Backlund","sequence":"first","affiliation":[{"name":"Biomedical Mass Spectrometry Facility, Intramural Research Program"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Henryk F.","family":"Urbanski","sequence":"additional","affiliation":[{"name":"Divisions of Neuroscience"},{"name":"Divisions of Reproductive and Developmental Sciences, Oregon National Primate Research Center, Beaverton, Oregon"},{"name":"Departments of Behavioral Neuroscience"},{"name":"Departments 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